Kinetic characterization of Escherichia coli outer membrane phospholipase A using mixed detergent-lipid micelles

Biochemistry ◽  
1989 ◽  
Vol 28 (3) ◽  
pp. 1139-1147 ◽  
Author(s):  
Anton J. G. Horrevoets ◽  
Tilman M. Hackeng ◽  
Hubertus M. Verheij ◽  
Ruud Dijkman ◽  
Gerard H. De Haas
Keyword(s):  
Escherichia Coli ◽  
Outer Membrane ◽  
Phospholipase A ◽  
Kinetic Characterization ◽  
Outer Membrane Phospholipase A ◽  
Lipid Micelles

scholarly journals Absence of the Outer Membrane Phospholipase A Suppresses the Temperature-Sensitive Phenotype of Escherichia coli degPMutants and Induces the Cpx and ςE Extracytoplasmic Stress Responses

2001 ◽  
Vol 183 (18) ◽  
pp. 5230-5238 ◽  
Author(s):  
Geoffrey R. Langen ◽  
Jill R. Harper ◽  
Thomas J. Silhavy ◽  
S. Peter Howard
Keyword(s):  
Escherichia Coli ◽  
Outer Membrane ◽  
Stress Responses ◽  
Elevated Temperatures ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Phospholipase A ◽  
Temperature Sensitive ◽  
Outer Membrane Phospholipase A ◽  
Temperature Sensitive Phenotype

ABSTRACT DegP is a periplasmic protease that is a member of both the ςE and Cpx extracytoplasmic stress regulons ofEscherichia coli and is essential for viability at temperatures above 42°C. [U-14C]acetate labeling experiments demonstrated that phospholipids were degraded indegP mutants at elevated temperatures. In addition, chloramphenicol acetyltransferase, β-lactamase, and β-galactosidase assays as well as sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis indicated that large amounts of cellular proteins are released from degP cells at the nonpermissive temperature. A mutation in pldA, which encodes outer membrane phospholipase A (OMPLA), was found to rescue degPcells from the temperature-sensitive phenotype. pldA degP mutants had a normal plating efficiency at 42°C, displayed increased viability at 44°C, showed no degradation of phospholipids, and released far lower amounts of cellular protein to culture supernatants. degP and pldA degP mutants containing chromosomal lacZ fusions to Cpx and ςE regulon promoters indicated that both regulons were activated in the pldA mutants. The overexpression of the envelope lipoprotein, NlpE, which induces the Cpx regulon, was also found to suppress the temperature-sensitive phenotype ofdegP mutants but did not prevent the degradation of phospholipids. These results suggest that the absence of OMPLA corrects the degP temperature-sensitive phenotype by inducing the Cpx and ςE regulons rather than by inactivating the phospholipase per se.

scholarly journals Bacteriocin Release Protein Triggers Dimerization of Outer Membrane Phospholipase A In Vivo

1999 ◽  
Vol 181 (10) ◽  
pp. 3281-3283 ◽  
Author(s):  
Niek Dekker ◽  
Jan Tommassen ◽  
Hubertus M. Verheij
Keyword(s):  
Escherichia Coli ◽  
Outer Membrane ◽  
Cell Envelope ◽  
Lag Time ◽  
Phospholipase A ◽  
Cross Linking ◽  
Outer Membrane Phospholipase A ◽  
Bacteriocin Release Protein ◽  
Chemical Cross Linking

ABSTRACT Bacteriocin release protein is known to activate outer membrane phospholipase A (OMPLA), which results in the release of colicin fromEscherichia coli. In vivo chemical cross-linking experiments revealed that the activation coincides with dimerization of OMPLA. Permeabilization of the cell envelope and dimerization were characterized by a lag time of 2 h.

scholarly journals Crystallization and preliminary X-ray analysis of outer membrane phospholipase A from Escherichia coli

FEBS Letters ◽  
1995 ◽  
Vol 373 (1) ◽  
pp. 10-12 ◽  
Author(s):  
Mieke Blaauw ◽  
Niek Dekker ◽  
Hubertus M. Verheij ◽  
Kor H. Kalk ◽  
Bauke W. Dijkstra
Keyword(s):  
Escherichia Coli ◽  
Outer Membrane ◽  
Phospholipase A ◽  
X Ray ◽  
Outer Membrane Phospholipase A

scholarly journals Molecular characterization of enterobacterial pldA genes encoding outer membrane phospholipase A.

1994 ◽  
Vol 176 (3) ◽  
pp. 861-870 ◽  
Author(s):  
R G Brok ◽  
E Brinkman ◽  
R van Boxtel ◽  
A C Bekkers ◽  
H M Verheij ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Molecular Characterization ◽  
Phospholipase A ◽  
Genes Encoding ◽  
Outer Membrane Phospholipase A

scholarly journals Assembly of Colicin A in the Outer Membrane of Producing Escherichia coli Cells Requires both Phospholipase A and One Porin, but Phospholipase A Is Sufficient for Secretion

2002 ◽  
Vol 184 (13) ◽  
pp. 3723-3733 ◽  
Author(s):  
Daniele Cavard
Keyword(s):  
Escherichia Coli ◽  
Monoclonal Antibodies ◽  
Outer Membrane ◽  
Polyclonal Antibodies ◽  
Sodium Dodecyl ◽  
Phospholipase A ◽  
Cell Fractionation ◽  
Heat Labile ◽  
Molecular Masses ◽  
Outer Membrane Phospholipase A

ABSTRACT Three oligomeric forms of colicin A with apparent molecular masses of about 95 to 98 kDa were detected on sodium dodecyl sulfate (SDS)-polyacrylamide gels loaded with unheated samples from colicin A-producing cells of Escherichia coli. These heat-labile forms, called colicins Au, were visualized both on immunoblots probed with monoclonal antibodies against colicin A and by radiolabeling. Cell fractionation studies show that these forms of colicin A were localized in the outer membrane whether or not the producing cells contained the cal gene, which encodes the colicin A lysis protein responsible for colicin A release in the medium. Pulse-chase experiments indicated that their assembly into the outer membrane, as measured by their heat modifiable migration in SDS gels, was an efficient process. Colicins Au were produced in various null mutant strains, each devoid of one major outer membrane protein, except in a mutant devoid of both OmpC and OmpF porins. In cells devoid of outer membrane phospholipase A (OMPLA), colicin A was not expressed. Colicins Au were detected on immunoblots of induced cells probed with either polyclonal antibodies to OmpF or monoclonal antibodies to OMPLA, indicating that they were associated with both OmpF and OMPLA. Similar heat-labile forms were obtained with various colicin A derivatives, demonstrating that the C-terminal domain of colicin A, but not the hydrophobic hairpin present in this domain, was involved in their formation.

scholarly journals A Conserved Histidine Residue of Escherichia Coli Outer-Membrane Phospholipase A is Important for Activity

1995 ◽  
Vol 234 (3) ◽  
pp. 934-938 ◽  
Author(s):  
Ronald G. P. M. Brok ◽  
Nick Dekker ◽  
Nathalie Gerrits ◽  
Hubertus M. Verheij ◽  
Jan Tommassen
Keyword(s):  
Escherichia Coli ◽  
Outer Membrane ◽  
Phospholipase A ◽  
Histidine Residue ◽  
Outer Membrane Phospholipase A

scholarly journals In Vitro Folding of Escherichia Coli Outer-Membrane Phospholipase A

1995 ◽  
Vol 232 (1) ◽  
pp. 214-219 ◽  
Author(s):  
Niek Dekker ◽  
Karin Merck ◽  
Jan Tommassen ◽  
Hubertus M. Verheij
Keyword(s):  
Escherichia Coli ◽  
Outer Membrane ◽  
Phospholipase A ◽  
Outer Membrane Phospholipase A
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